RENAL SINGLE-NUCLEAR TRANSCRIPTOMICS IDENTIFIES NOVEL THERAPEUTIC TARGETS IN A PRECLINICAL MODEL OF CHRONIC KIDNEY DISEASE.

IF 3 Q1 UROLOGY & NEPHROLOGY
Kidney360 Pub Date : 2025-08-08 DOI:10.34067/KID.0000000945
Alejandro R Chade, Sathesh K Sivasankaran, Rhys Sitz, Elizabeth A McCarthy, Alfonso Eirin
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引用次数: 0

Abstract

Background and hypothesis: The prevalence of chronic kidney disease (CKD) is on the rise, and precision strategies to offset the progression to end-stage kidney disease are needed. We used a well-established pre-clinical translational model of CKD in swine and single nucleus RNA sequencing (snRNA-seq) to identify potential therapeutic targets via characterization of the renal cell-specific transcriptomic landscape.

Methods: Normal and CKD pigs were studied in vivo and ex vivo after 14 weeks (n=6/group). In randomly selected pigs (n=3/group), kidneys were harvested, nuclei isolated, libraries prepared, and snRNA-seq performed. Protein expression of candidate differentially expressed genes (DEGs, log2FC>0.25, adjusted p-value<0.05) was determined by immunohistochemistry, and their expression in primary normal and CKD renal vascular endothelial cells (RECs) was modulated (siRNA) in vitro.

Results: A total of 52,213 nuclei were analyzed. Thirty clusters were identified and filtered by canonical gene markers, revealing 16 unique renal cell types. Endothelial cells were the top cell type exhibiting the highest number of DEGs; which were subsequently filtered by angiogenesis-, inflammation-, and fibrosis (major injurious pathways altered in CKD kidneys). Venn diagram analysis identified 5 unique overlapping DEGs in endothelial cells: VWF, LAMA3, and KDR upregulated, and PTGIS and ICAM1 downregulated in CKD versus normal kidneys. Venn diagram analysis indicates that VWF, LAMA3, and ICAM1 participate in inflammatory and fibrotic signaling. Renal protein expression of these DEGs matched snRNA-seq findings. Furthermore, in vitro silencing of VWF and LAMA3 ameliorated endothelial cell inflammatory and fibrotic signaling.

Conclusions: Our work characterized the single-nuclear renal cell transcriptomic landscape of a translational model of CKD and singled out genes implicated in major renal injury pathways. These genes could serve as potential targets to pave the way for new therapeutic strategies in patients with CKD.

肾脏单核转录组学在慢性肾脏疾病的临床前模型中确定了新的治疗靶点。
背景与假设:慢性肾脏疾病(CKD)的患病率正在上升,需要精确的策略来抵消进展到终末期肾脏疾病。我们使用了一个完善的猪CKD临床前转化模型和单核RNA测序(snRNA-seq),通过表征肾细胞特异性转录组学景观来确定潜在的治疗靶点。方法:正常猪和CKD猪14周后在体内和离体实验(n=6/组)。在随机选择的猪(n=3/组)中,取肾,分离细胞核,制备文库,并进行snrna测序。候选差异表达基因(DEGs, log2FC>0.25,调整p值)的蛋白表达结果:共分析了52,213个细胞核。通过典型基因标记对30个簇进行鉴定和筛选,发现16种独特的肾细胞类型。内皮细胞是DEGs数量最多的细胞类型;随后被血管生成、炎症和纤维化(CKD肾脏的主要损伤途径改变)过滤。维恩图分析发现内皮细胞中有5个独特的重叠deg:与正常肾脏相比,CKD中VWF、LAMA3和KDR上调,PTGIS和ICAM1下调。维恩图分析表明VWF、LAMA3和ICAM1参与炎症和纤维化信号传导。这些deg的肾蛋白表达与snRNA-seq结果相符。此外,体外沉默VWF和LAMA3可改善内皮细胞炎症和纤维化信号。结论:我们的工作表征了CKD翻译模型的单核肾细胞转录组景观,并挑出了与主要肾损伤途径相关的基因。这些基因可以作为潜在的靶点,为CKD患者的新治疗策略铺平道路。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Kidney360
Kidney360 UROLOGY & NEPHROLOGY-
CiteScore
3.90
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0.00%
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